Odorous emissions of synthetic turf and its relationship with local communities.

Synthetic turf has been a mainstay of field sports and local communities for decades, and in that time, has faced both community and government pressure to ensure its safety and fitness for purpose. Considerable research and regulations have been applied to synthetic turf with regards to its safety, construction, potential toxicity, sports impact, as well as environmental considerations. However, very little attention has been paid to reports of odorous impacts from synthetic turf fields. This is problematic as odours are both a source of most complaints by communities towards other industries, as well as the fact that synthetic turf has a unique placement within communities themselves. It is wholly possible that the concerns surrounding synthetic turf are being modulated by the odours that the fields themselves produce through previously identified psychological mechanisms. As a result, ensuring good standards for synthetic turf with regards to odorous emissions should be benchmarked for community acceptability. This review investigates prior research into synthetic turf with regards to identified volatile organic compounds emitted, as well as proposing the means by which community stakeholders engage with synthetic turf, as well as how they should be consulted. From here, this review provides trajectories for future research within this space, and how regulatory bodies should address potential issues. This research space is currently in its infancy and therefore information relating to synthetic turf odour factors must be carefully considered.

Author Correction: Anak Krakatau triggers volcanic freezer in the upper troposphere.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Anak Krakatau triggers volcanic freezer in the upper troposphere.

Volcanic activity occurring in tropical moist atmospheres can promote deep convection and trigger volcanic thunderstorms. These phenomena, however, are rarely observed to last continuously for more than a day and so insights into the dynamics, microphysics and electrification processes are limited. Here we present a multidisciplinary study on an extreme case, where volcanically-triggered deep convection lasted for six days. We show that this unprecedented event was caused and sustained by phreatomagmatic activity at Anak Krakatau volcano, Indonesia during 22-28 December 2018. Our modelling suggests an ice mass flow rate of ~5 × 106 kg/s for the initial explosive eruption associated with a flank collapse. Following the flank collapse, a deep convective cloud column formed over the volcano and acted as a 'volcanic freezer' containing ~3 × 109 kg of ice on average with maxima reaching ~1010 kg. Our satellite analyses reveal that the convective anvil cloud, reaching 16-18 km above sea level, was ice-rich and ash-poor. Cloud-top temperatures hovered around -80 °C and ice particles produced in the anvil were notably small (effective radii ~20 µm). Our analyses indicate that vigorous updrafts (>50 m/s) and prodigious ice production explain the impressive number of lightning flashes (~100,000) recorded near the volcano from 22 to 28 December 2018. Our results, together with the unique dataset we have compiled, show that lightning flash rates were strongly correlated (R = 0.77) with satellite-derived plume heights for this event.

Follicular dynamics, circulating progesterone, and fertility in Holstein cows synchronized with reused intravaginal progesterone implants that were sanitized by autoclave or chemical disinfection.

This experiment aimed to compare circulating progesterone (P4), follicular dynamics, and fertility during reuse of intravaginal P4 implants that were sanitized by autoclave or chemical disinfection in lactating Holstein cows submitted to fixed-time artificial insemination (FTAI). For this, 123 primiparous and 226 multiparous cows from 2 farms, averaging (mean ± standard deviation) 163.9 ± 141.9 d in milk, 35.7 ± 11.3 kg of milk/d, and a body condition score of 2.9 ± 0.5, were enrolled in the study. Cows were randomly assigned to 1 of 2 treatments using a completely randomized design and each cow received a reused implant (1.9 g of P4; previously used for 8 d) that was either autoclaved (AUT; n = 177) or chemically disinfected (CHEM; n = 172) on d -10. Also on d -10, cows received 2 mg of estradiol benzoate and 100 µg of GnRH. On d -3, cows received 25 mg of dinoprost (PGF2α). A second PGF2α was given on d -2, along with 1 mg of estradiol cypionate and P4 implant removal. Cows received FTAI on d 0. A subset of cows (n = 143) was evaluated by ultrasound on d -10, -8, -6, -3, -2, 0, and 5 to identify ovarian structures, and blood was sampled on d -10, -3, and -2 for P4 concentrations by RIA. Pregnancy diagnoses were performed at d 32 and 60. Statistical analyses was performed using PROC-MIXED for continuous variables and PROC-GLIMMIX of SAS 9.4 (SAS Institute Inc., Cary, NC) for binomial variables. The treatments did not differ in circulating P4 on d -10 or -3, but P4 was greater on d -2 in CHEM cows. Ovulation to the treatments on d -10 was associated with lower circulating P4 on d -10 (2.0 vs. 3.1 ng/mL) and resulted in greater P4 on d -3 (4.0 vs. 2.4 ng/mL) and more cows with a corpus luteum on d -3 (100 vs. 40%) than nonovulating cows. Cows that ovulated to d -10 treatments were more likely to have a synchronized new follicular wave (97.9 vs. 63.2%) and had an earlier wave emergence (1.9 vs. 2.6 d), resulting in less cows ovulating a persistent follicle (0.0 vs. 35.7%). Type of P4 implant, corpus luteum presence on d -10, and ovulation to d -10 treatments did not affect fertility (pregnancy per AI; P/AI). However, P/AI on farm A was greater than on farm B at 32 (40.8 vs. 27.8%) and 60 d (35.8 vs. 24.3%), independent of treatment. In conclusion, P4 implants with different P4 release patterns did not produce detectable differences in follicular dynamics, synchronization rate, or P/AI. Nevertheless, presence of corpus luteum or ovulation at the beginning of the FTAI protocol affected reproductive variables, such as timing and synchronization of follicular wave emergence, and size of the ovulatory follicle. Beyond that, more overall synchronized cows became pregnant to the FTAI protocol.

Artificial cloud test confirms volcanic ash detection using infrared spectral imaging.

Airborne volcanic ash particles are a known hazard to aviation. Currently, there are no means available to detect ash in flight as the particles are too fine (radii < 30 µm) for on-board radar detection and, even in good visibility, ash clouds are difficult or impossible to detect by eye. The economic cost and societal impact of the April/May 2010 Icelandic eruption of Eyjafjallajökull generated renewed interest in finding ways to identify airborne volcanic ash in order to keep airspace open and avoid aircraft groundings. We have designed and built a bi-spectral, fast-sampling, uncooled infrared camera device (AVOID) to examine its ability to detect volcanic ash from commercial jet aircraft at distances of more than 50 km ahead. Here we report results of an experiment conducted over the Atlantic Ocean, off the coast of France, confirming the ability of the device to detect and quantify volcanic ash in an artificial ash cloud created by dispersal of volcanic ash from a second aircraft. A third aircraft was used to measure the ash in situ using optical particle counters. The cloud was composed of very fine ash (mean radii ~10 µm) collected from Iceland immediately after the Eyjafjallajökull eruption and had a vertical thickness of ~200 m, a width of ~2 km and length of between 2 and 12 km. Concentrations of ~200 µg m(-3) were identified by AVOID at distances from ~20 km to ~70 km. For the first time, airborne remote detection of volcanic ash has been successfully demonstrated from a long-range flight test aircraft.

Pulmonary ventilation/perfusion single photon emission tomography--Initial experience of a Nuclear Medicine Department.

INTRODUCTION: Lung ventilation/perfusion scintigraphy with planar images (V/QS-planar) is very useful for the diagnosis and follow-up of pulmonary thromboembolism (PTE). Acquiring tomographic images (V/QS-SPECT) is a recent development with potential to increase the technique's accuracy. The purpose of this work is to evaluate the added benefits of V/QS-SPECT studies as opposed to traditional planar imaging. PATIENTS AND METHODS: We prospectively revised 53 V/QS-planar and V/QS-SPECT exams, performed according to the European Association of Nuclear Medicine guidelines. We evaluated the exams independently, by consensus of two Nuclear Medicine physicians. For both methods, we gave each lung a score expressing the dimension and extension of perfusion defects with normal ventilation. For each lung, we compared the scores with the paired Wilcoxon test, estimating the 95% confidence interval (95 CI) for the respective difference. RESULTS: We performed V/QS-SPECT exams without technical difficulties. The paired Wilcoxon test estimated the score difference to be -0.75 (95 CI of -1.0 to -0.5; p-value=9.6 × 10(-7)), expressing a statistically significant difference of about 1 subsegmental defect between both methods, with V/QS-SPECT detecting more defects. DISCUSSION: The results demonstrate that V/QS-SPECT identifies a slightly larger number of perfusion defects than V/QS-planar, suggesting a higher sensitivity of this technique. However, more studies are necessary to evaluate the clinical meaning of this fact. CONCLUSION: V/QS-SPECT demonstrates a higher capability to identify perfusion defects. This method looks promising, allowing for a greater role of this exam in pulmonary thromboembolism diagnosis and follow-up.

Increasing estradiol benzoate, pretreatment with gonadotropin-releasing hormone, and impediments for successful estradiol-based fixed-time artificial insemination protocols in dairy cattle.

With the objective to optimize fixed-time artificial insemination (FTAI) protocols based on estradiol benzoate (EB) and progesterone (P4), we performed 2 experiments (Exp.) in dairy cows. In Exp. 1 (n=44), we hypothesized that increased EB (EB3=3 mg vs. EB2=2 mg) on d 0 would improve synchronization of ovarian follicle wave emergence. Likewise, in Exp. 2 (n=82), we hypothesized that a GnRH treatment on d -3 (early in a follicular wave on d 0) versus d -7 (presence of a dominant follicle on d 0) would better synchronize wave emergence. Moreover, results from both experiments were combined to identify reasons for the lack of synchronization. All cows were treated with EB at the time of introduction of a P4 implant (d 0). On d 7, cows were given 25 mg of prostaglandin F2α; on d 8, the implant was removed and cows were given 1mg of estradiol cypionate. All cows received FTAI on d 10. In both experiments, daily ultrasound evaluations were performed and, in Exp. 2, circulating P4 was evaluated during the protocol. Pregnancy per artificial insemination (P/AI) was determined on d 31 and 59 after FTAI. In Exp. 1, EB dose did not change time to wave emergence, but EB3 compared with EB2 decreased the percentage of cows with a corpus luteum on d 7 (19.8 vs. 55.3%) and time to ovulation (10.4 vs. 10.9 d). In Exp. 2, although we detected a tendency for delayed follicle wave emergence after the start of the FTAI protocol in cows ovulating to GnRH given on d -7, there was no difference in percentage of cows with a synchronized wave emergence (~80%). Regardless of treatment, more cows with P4<0.1 ng/mL, compared with P4≥0.1 and <0.22 ng/mL at the time of AI, ovulated to the protocol (81.2 vs. 58.0%) and had increased P/AI (47.4 vs. 21.4%). An analysis of data from both experiments showed that only 73.8% (93/126) of cows had synchronized wave emergence, and only 77.8% (98/126) of cows ovulated at the end of the protocol. Fertility was much greater in cows that had emergence of a new wave synchronized and ovulated to end of the protocol [P/AI 61.3% (46/75)] compared with cows that failed to present one or both of the outcomes above [15.7% (8/51)]. Thus, although current FTAI protocols using EB and P4 produce P/AI between 30 and 40% for lactating dairy cows, there remains room for improvement because less than 60% (75/126) of the cows were correctly synchronized. Starting the FTAI protocol without the dominant follicle or increasing the dose of EB to 3mg was not effective in increasing synchronization rate.

Effect of injectable vitamin E on incidence of retained fetal membranes and reproductive performance of dairy cows.

The objectives were to evaluate the effects of injectable vitamin E during the last 3 wk prepartum on the incidence of retained fetal membranes (RFM) and reproductive performance. Dairy cows (n=890), 390 Holsteins (132 nulliparous and 258 parous) and 500 crossbred Holstein × Gyr (199 nulliparous and 301 parous), from 3 dairy farms in Brazil were assigned to the study. In all 3 farms, from October to March, prepartum cows grazed tropical grasses and received 2 kg/d of a mixture of finely ground corn, soybean meal, and minerals and vitamins. From April to September prepartum cows received a total mixed ration composed of corn silage, finely ground corn, soybean meal, and minerals and vitamins. During the prepartum period, cows were fed 280 (farm 1), 390 (farm 2), and 480 IU (farm 3) of supplemental vitamin E per day, and throughout postpartum, cows were fed 370 (farm 1), 500 (farm 2), and 600 (farm 3) IU of supplemental vitamin E. Within each farm, cows were randomly assigned to remain as untreated controls or to receive 3 i.m. injections of 1,000 IU each of dl-α-tocopherol administered at 19.2 ± 4.3, 12.9 ± 3.3, and 6.2 ± 2.9 d before calving (VitE). Blood was sampled from 141 cows immediately before enrollment to determine the α-tocopherol and cholesterol statuses. Blood was also sampled and analyzed for concentrations of cortisol and nonesterified fatty acids in the last 3 wk of gestation. The serum concentration of α-tocopherol or α-tocopherol:cholesterol ratio did not differ between treatments and averaged 2.97 ± 0.10 µg/mL and 4.46 ± 0.16 × 10(-3), respectively. In total, 53.2% of the cows had an inadequate concentration of serum α-tocopherol based on the 3.0 µg/mL cut-off for adequacy. The risk of RFM decreased as serum α-tocopherol increased. Milk production did not differ between controls and VitE cows. Treatment with injectable α-tocopherol decreased RFM from 20.1 to 13.5%, decreased incidence of stillbirth from 14.9 to 6.8%, and tended to decrease death by 200 d postpartum. VitE cows tended to have improved pregnancy per insemination at first AI (36.7 vs. 30.1%) because of decreased pregnancy loss from 31 to 62 d of gestation (12.5 vs. 20.5%). Despite a similar insemination rate, VitE cows had 22% greater pregnancy rate than control cows. Cows receiving vitamin E had decreased circulating cortisol and nonesterified fatty acids around calving. In summary, when cows were fed limited amounts of supplemental vitamin E, 28 to 48% of the recommendations, prepartum supplementation with injectable α-tocopherol decreased incidence of RFM and improved reproduction.

Production of microparticles with gelatin and chitosan.

In the past few decades, the textile industry has significantly increased investment in research to develop functional fabrics, with a special focus on those aggregating values. Such fabrics can exploit microparticles inferior to 100 µm, such as those made by complex coacervation in their creation. The antimicrobial properties of chitosan can be attributed to these microparticles. Developing particles with uniform structure and properties would facilitate the control for the eventual release of the core material. Thus, a complex coacervation between gelatin and chitosan was studied, and the optimal conditions were replicated in the encapsulation of limonene. Spherical particles formed had an average diameter (D3,2) of 30 µm and were prepared with 89.7% efficiency. Cross-linking of these microparticles using glutaraldehyde and tripolyphosphate was carried out before spray drying. After drying, microparticles cross-linked with glutaraldehyde were oxidized and clustered and those that were cross-linked with tripolyphosphate resisted drying and presented a high yield.

Molecular characterization of bromeliads from northeast Brazil.

Bromeliaceae is an important botany family that includes many species with economic value; demand for members of this family is increasing. However, illegal collection frequently occurs, drastically reducing the species populations; thus, it is necessary to collect and store Bromeliaceae genetic material. In this study, we identified and quantified genetic variability of the Bromeliad family using dominant markers to create the first Germplasm Bank in the northeast region of Brazil. Molecular tools were used to characterize the collected accessions. The combination of 11 inter-simple sequence repeats and 13 random amplified polymorphic DNA markers were used to detect the genetic variability of wild bromeliad accessions.

Antinociceptive, anti-inflammatory and antioxidant activities of aqueous extract from Remirea maritima (Cyperaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Remirea maritima Aubl., popularly known as "capim-da-praia", is popularly employed in the treatment of diarrhea, kidney disease, fever, and for analgesic and anti-inflammatory purposes through the preparation of teas. Few studies have focused on the chemical composition and its biological properties. AIM OF THE STUDY: This work evaluated the antinocipetive, anti-inflammatory and antioxidant activities of the aqueous extract from Remirea maritima Aubl. as well as the isolation and identification of the chemical compounds. MATERIALS AND METHODS: Compounds were isolated from aqueous extract of Remirea maritima through preparative HPLC and the structures were identified by means of NMR and MS analysis. The tests for antinociceptive, anti-inflammatory, and antioxidant activities, along with motor coordination test (Rota rod), were performed over the aqueous extract. RESULTS: The phytochemical investigation of aqueous extract of Remirea maritima resulted in the isolation of three flavone glycosides. The structures of these compounds were determined by means of MS and 1D and 2D NMR data as vitexin-2″-O-ß-D-glucopyranoside, isovitexin-2″-O-ß-D-glucopyranoside, and luteolin-7-O-glucuronide. Acute pretreatment with aqueous extract (100, 200 or 400mg/kg, i.p.) caused a significant decrease (p<0.001) in the number of abdominal writhes. In the formalin test, higher doses significantly inhibited the late (inflammatory pain) phase of formalin-induced licking (p<0.05 or 0.001). In the hot plate test, there was no significant difference in nociceptive behavior, discarding the possible central effect of the aqueous extract. In the rota rod test, it was verified that the aqueous extract in all concentration evaluated does not alter the motor coordination of mice, such antinociceptive results were unlikely to be caused by motor abnormality. In the peritonitis test, induced by carrageenan, the treatment with aqueous extract produced a significant reduction in leukocyte migration in all concentration evaluated. Additionally, a significant reduction of lipoperoxidation (TBARS test) and in nitric oxide formation (.NO Scavenging assay) was observed in antioxidant activity assay. CONCLUSION: The biological and phytochemical investigations of the aqueous extract of Remirea maritima resulted in the identification of three flavone glycosides that have been described here for the first time in Remirea and effective analgesic activity in various pain models, probably mediated via the inhibition of peripheral mediators which could be related to its strong antioxidant effect observed in vitro.

Heart rate variability in chronic Chagas patients before and after treatment with benznidazole.

BACKGROUND: There is no consensus regarding large-scale use of benznidazole to treat Chagas disease, because of its toxicity and low efficacy during the chronic phase. OBJECTIVES: To evaluate heart rate variability in chronic Chagas patients before and after treatment with benznidazole. METHODS: Twenty-one Chagas patients with positive blood cultures and/or PCR received benznidazole (5 mg/kg twice daily for 60 days) and were matched for age and gender with 24 Chagas individuals with negative blood cultures, as controls. R-R intervals were assessed in time and frequency domains using an autoregressive algorithm under three different conditions: baseline, cold face and active tilt tests. Power spectral densities in low and high-frequency bands were estimated in absolute and normalized units. Data were expressed as mean±SD or medians (lower and upper quartiles). Groups were compared using non-paired or paired Student's T, Mann-Whitney or Wilcoxon tests, as required. The significance level was 5%. RESULTS: The groups were comparable regarding age, gender and clinical disease forms. There were no differences in temporal and spectral indices between groups in baseline and cold face tests. Treated patients presented significantly lower tachycardic responses in mean R-R intervals during the active tilt test (p=0.0200) than controls did. Genetic characterization of T. cruzi isolates from treated group samples showed that 100% belonged to genotype II. CONCLUSION: For the first time in the medical literature, we detected sympathetic impairment during the active tilt test in chronic Chagas patients treated with benznidazole. This finding may be partially explained by benznidazole neurotoxicity.

Assessment of salivary gland function in Sjögren's syndrome: the role of salivary gland scintigraphy.

Salivary gland scintigraphy (SGS) is a non invasive method of salivary gland function assessment. This technique is easy to perform, reproducible and well tolerated by patients. Additionally, an abnormal salivary gland scintigraphy result is accepted by the American-European consensus group as a criterion for the diagnosis of Sjögren's syndrome. Scintigraphic evaluation of salivary gland function also plays an important role in therapeutic decision and patient follow-up. Schall's categorical classification is usually considered the standard method for salivary scintigraphy interpretation, though subjective and with limited capacity to discriminate borderline results. In order to improve the diagnostic accuracy of SGS, there has been an increasing interest in the quantification of glandular function. However, the debate on the most reliable and suitable parameters for the diagnosis of SS persists.

[Salivary gland scintigraphy in the evaluation of patients with sicca complaints]. / Cintigrafia das glândulas salivares major na avaliação de doentes com queixas secas.

UNLABELLED: Salivary gland scintigraphy is a non invasive method widely accepted as an objective assessment of salivary gland function and a diagnostic criterion of primary Sjögren's Syndrome. In this study we evaluated the performance of qualitative and semi--qualitative salivary gland scintigraphy in primary Sjögren's Syndrome. Additionally we aimed to identify the scintigraphic patterns of salivary gland involvement in this condition. PATIENTS AND METHODS: Observational study of fifty--six women with suspected primary Sjögren's Syndrome (pSS). Investigation included clinical history and observation, ophtalmologic examination, autoantibodies (SSA and SSB) determination, minor salivary gland biopsy and salivary gland scintigraphy interpreted according to Schall classification, visual quality of radioactivity uptake, morphology of time-activity curves and semi-quantitative parameters of uptake and excretion. RESULTS: Twenty patients fulfilled diagnostic criteria for pSS. Submandibular glands showed more pronounced functional impairment. In pSS patients, all scintigraphic parameters were significantly lower on these glands: a) qualitative evaluation ' visual quality of uptake (p=0,003), morphology of time-activity curves (p=0,001) and Schall classification (p<0,001); b) semi-quantitative parameters ' maximum counts (p=0,005), maximum counts/second/MBq administered (p=0,01), index of maximum counts versus counts at first minute (p=0,002) and excretion index (p=0,006). CONCLUSIONS: Despite being qualitative and observer-dependent, Schall classification is valuable for the diagnosis of primary Sjögren's Syndrome. Nevertheless, semi-quantitative evaluation of salivary gland scintigraphy reveals significant differences in pSS patients and may have incremental value for the interpretation of salivary gland scintigraphy results.

Kinetics of Bacillus thuringiensis var. israelensis growth on high glucose concentrations.

The kinetic and general growth features of Bacillus thuringiensis var. israelensis were evaluated. Initial glucose concentration (S0) in fermentation media varied from 10 to 152 g/l. The results afforded to characterize four morphologically and physiologically well-defined culture phases, independent of S0 values: Phase I, vegetative growth; Phase II, transition to sporulation; Phase III, sporulation; and Phase IV, spores maturation and cell lysis. Important process parameters were also determined. The maximum specific growth rates (microX,m) were not affected with S0 up to 75 g/l (1.0-1.1 per hour), but higher glucose concentrations resulted in growth inhibition by substrate, revealed by a reduction in microX,m values. These higher S0 values led to longer Phases III and IV and delayed sporulation. Similar biomass concentrations (Xm=15.2-15.9 g/l) were achieved with S0 over 30.8 g/l, with increasing residual substrate, suggesting a limitation in some other nutrients and the use of glucose to form other metabolites. In this case, with S0 from 30.8 to 152 g/l, cell yield (YX/S) decreased from 0.58 to 0.41 g/g. On the other hand, with S0=10 g/l growth was limited by substrate, and YX/S has shown its maximum value (0.83 g/g).

Mixed inflammatory/regulatory cytokine profile marked by simultaneous raise of interferon-gamma and interleukin-10 and low frequency of tumour necrosis factor-alpha(+) monocytes are hallmarks of active human visceral Leishmaniasis due to Leishmania chagasi infection.

Considering the complexity of the immunological events triggered during active visceral Leishmaniasis (VL), the relevance of the segregation of the immune response during human VL into type 1 and type 2 still remains unclear. For this purpose, in individuals living in risk areas for VL, we have evaluated especially asymptomatic individuals and patients with active VL, the plasmatic levels of cytokines and reactive nitrogen species under ex vivo conditions. In addition, we have also performed an analysis of intracellular cytokine patterns of circulating leucocytes after short-term culture, particularly in the absence of antigenic-specific stimulation, in order to reflect dynamic events of immune response in vivo during Leishmania chagasi infection. Although asymptomatic individuals and non-infected subjects presented a similar immunological profile, an outstanding inflammatory/regulatory profile, based on higher plasmatic levels of cytokines such as interleukin (IL)-8, interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, IL-6 and IL-10, was associated with clinical status observed in active VL. In this context, we hypothesize that IL-10, through its ability to inhibit anti-leishmanial macrophage activation, associated with the lower frequency of TNF-alpha(+) monocytes and ordinary levels of nitrite and nitrate are the major mechanisms associated with disease onset.

Cystoisospora belli: in vitro multiplication in mammalian cells.

Intracellular development of Cystoisospora belli was demonstrated in 4 different mammalian cell lines. Human ileocecal adenocarcinoma (HCT-8), epithelial carcinoma of lung (A549), Madin-Darby bovine kidney (MDBK), and African green monkey kidney (VERO) were exposed in vitro to C. belli sporozoites, which had been isolated from the feces of HIV-AIDS patients. Parasites invaded all the cellular types between 4 and 12h after exposure and multiplication was demonstrated after 24 h. Grater number of merozoites formed in VERO cells, followed by HCT-8. In the MDBK and HCT-8 cells, the parasitophorous vacuole was less evident and immobile merozoites were observed in the cytoplasm. In VERO cells, one or several parasitophorous vacuoles contained up to 16 mobile sporozoites. No oocysts were found in any of the cell types used. VERO cells may be suitable for studies of the interaction between parasite and host cells.

Immune response in human visceral leishmaniasis: analysis of the correlation between innate immunity cytokine profile and disease outcome.

We investigated the cytokine profile of cells of the innate immune response and its association with active (ACT), asymptomatic (AS) and cured (CUR) human visceral leishmaniasis (VL), as well as noninfected (NI) subjects. The frequency of cytokine-producing cells was determined after short-term in vitro incubation of whole peripheral blood samples with soluble Leishmania antigen (SLA). Our data demonstrated a predominant type 2 cytokine profile in NI and ACT. In NI, we observed an increase of IL-4+ neutrophils, IL-10+ eosinophils besides a decrease of tumour necrosis factor (TNF)-alpha+ eosinophils/monocytes. Yet in ACT, we observed an increase of IL-4+ neutrophils and natural killer (NK) cells and IL-10+ monocytes, a reduced frequency of IL-12+ and IFN-gamma+ eosinophils and lower levels of TNF-alpha+ and IL-12+ monocytes. AS presented a mixed profile, characterized by an increase of IFN-gamma+ neutrophils/eosinophils and NK cells, of IL-12+ eosinophils/monocytes, as well as increase of IL-4+ neutrophils and NK cells and IL-10+ eosinophils/monocytes. In contrast, CUR was characterized by a type 1 response with an increase of IFN-gamma+ neutrophils/eosinophils and NK cells, associated with an increase in IL-12+ monocytes. In conclusion, we show a correlation between innate immune cytokine patterns and clinical status of VL, suggesting that these cells, in addition to other factors, may contribute to the cytokine microenvironment in which Leishmania-specific T cells are primed and to disease outcome.